1982-2012年江苏省审定油菜品种系谱及性状分析

分析了1982-2012年江苏省审定的62个油菜品种的亲本系谱、品种特性以及主要农艺性状和经济性状特点。结果表明,江苏省审定的甘蓝型油菜品种的原始亲本为胜利油菜,低芥酸性状主要源自Zephyr、Primor、Midas和荣山等,低硫苷性状主要源自Tower、Wesroona、Marnoo和Wesbrook等。江苏省审定的油菜品种的主要农艺性状中,生育期逐步缩短,并基本稳定在243 d左右;株高变化较大,因品种而异。4个产量(产油量)构成因素中,含油量年平均增幅最大,尤其是在2003-2012年间,年平均增幅达到0.65%;单株角果数和每角粒数有上升趋势,而千粒重则变化相对较小。受双低性状对产量性状的负效应影响,产量及产油量在1982-2012年间的年平均增幅均不大,但优质常规品种的产量与产油量的年平均增幅较大,分别为23.10 kg/hm2和13.20 kg/hm2。研究结果可为江苏省油菜新品种的选育与推广提供参考。     

Identification of antibiosis and tolerance in rice varieties carrying brown planthopper resistance genes

Brown planthopper (BPH) [Nilaparvata lugens (Stål) (Hemiptera: Delphacidae)] is a major pest in rice [Oryza sativa L. (Poaceae)] production. Identification of resistance genes and development of BPH‐resistant varieties is an economical and effective way to control this pest. In this study, BPH honeydew excretion, survival rate, and emergence rate were used as indicators to detect the antibiotic level, whereas the relative growth rates of plant height (R_H) and fresh weight (R_W), and the number of days until yellowing were used to identify the level of tolerance to BPH in rice varieties. Rice varieties Swarnalata and B5, which showed high levels of antibiosis and tolerance to BPH, thus were highly resistant in the seedling bulk test; Mudgo and T12, which showed moderate resistance to the insects, had a high level of tolerance and moderate antibiosis to BPH. Varieties Rathu Heenati, ARC 10550, and Chin Saba were identified to be susceptible to BPH, showing a moderate level of tolerance and no antibiosis. In comparison to the evaluation methods of BPH resistance, the honeydew excretion and survival rate could be used to detect the antibiotic level, and the R_H, R_W, or leaf yellowing days could be employed as indicators to evaluate the rice varieties’ tolerance. Overall, a combined application of these indicators can effectively identify the levels of antibiosis and tolerance to BPH in rice varieties, and BPH‐resistance levels of the varieties were mainly determined by the antibiosis level. The results should help in understanding BPH‐resistance categories of rice varieties and for resistance breeding.     

Identification and analysis of whole microcystin synthetase genes from two Korean strains of the cyanobacterium Microcystis aeruginosa

Microcystins are cyanobacterial hepatotoxins, and are produced by nonribosomal enzyme complexes, mcy gene cluster. In this study, we report on whole mcy gene clusters from two Korean strains of M. aeruginosa that were blooming in Lake Paldang (FCY-26) and Geum river (FCY-28). Their specific gene locus, amino acid information, and sub-cluster orientation were also characterized in both strains. Both gene clusters are of 55 kb, and also each length, number and the arrangement are identical. Their sequence analysis revealed a cluster of 10 genes (mcyA, B, C, D, E, F, G, H, I, and J) involved in the biosynthesis of microcystin, and mcyABC and mcyDEFGHIJ formed two polycistronic operon structures that are transcribed bidirectionally from a central promoter region between mcyA and mcyD. The analysis of SNPs provided different nucleotide composition and amino acid variations in two Korean strains of M. aeruginosa. This approach is useful to develop genetic indicators identifying toxic cyanobacteria and their cyanotoxins, and helpful for a better understanding of the diversities of mcy gene clusters, the biosynthesis of microcystin, and the mediation of environmental parameters causing algal blooming and HABs.     

Identification and analysis of bacterial virulence genes in vivo

Signature-tagged mutagenesis is a mutation-based screening method for the identification of virulence genes of microbial pathogens. Genes isolated by this approach fall into three classes: those with known biochemical function, those of suspected function and some whose functions cannot be predicted from database searches. A variety of in vitro and in vivo methods are available to elucidate the function of genes of the second and third classes. We describe the use of some of these approaches to study the function of the Salmonella pathogenicity island 2 type III secretion system of Salmonella typhimurium. This virulence determinant is required for intracellular survival. Secretion by this system is induced by an acidic pH, and its function may be to alter trafficking of the Salmonella-containing vacuole. Use of a temperature-sensitive non-replicating plasmid and competitive index tests with other genes show that in vivo phenotypes do not always correspond to those predicted from in vitro studies.     

Identification and functional analysis of LsMNPV anti-apoptosis genes

Three anti-apoptosis genes, Ls-iap2, iap3 and p49 were found in Leucania separata multiple nuclear polyhedrovirus. Amino acid sequence homology of Ls-IAP2 and Ls-IAP3 with Op-IAP2 and Op-IAP3 from Orgyia pseddotsugata MNPV were 20% and 42%, while that of Ls-P49 is 28% with Sl-P49 from Spodoptera littorolis MNPV. Ls-IAP2 contains one baculoviral IAP repeat (BIR) domain followed by a RING domain, while Ls-IAP3 contains two BIRs and a RING. Ls-P49 contains a reactive site loop, predicted cleavage site (KKLD(74) downward arrow G) that is different from Sl-P49 (TVID(94) downward arrow G). Expressed Ls-iap3 or Ls-p49 under presence of actinomycin D in SF9 cells, DNA ladder assay revealed that Ls- IAP3 or Ls-P49 could block the apoptosis of SF9 cells induced by actinomycin D. Replication of p35 deficient-mutant Autographa californica MNPV in SF9 cells was also rescued when Ls-iap3 or Ls-p49 was expressed transiently. No anti-apoptotic activity was observed for Ls-IAP2. The results showed that both of Ls-IAP3 and Ls-P49 were functional apoptotic suppressors in SF9 cells.     

Investigation of the origin and transmission of linear mitochondrial plasmid based on phylogenetic analysis in Japanese rapeseed varieties.

A linear mitochondrial plasmid is present in some varieties of rapeseed. To elucidate its origin and transmission the author investigated types of mitochondrial genome and the presence of plasmid in 78 rapeseed varieties and landraces in Japan and carried out a comparative analysis using the breeding history of Japanese rapeseed varieties. The mitochondrial genome of rapeseed was classified roughly into 2 types, type I (nap) and type II (cam). Type II rapeseed mitochondria closely resembles that of Brassica rapa, which is a related species of rapeseed. In this study, the author found that all varieties with type II mitochondria originated from interspecific crosses between rapeseed (B. napus) and B. rapa. This indicates that type II cytoplasm was introduced to rapeseed through a breeding program. The presence of plasmid was limited to B. rapa landraces and rapeseed varieties that arose by interspecific crosses between B. napus and B. rapa. The results suggest that mitochondrial plasmid is of B. rapa origin and that it has been introduced into rapeseed by interspecific crosses in a modern breeding program, as in the case of the mitochondrial genome. Phylogenetic study of Japanese rapeseed varieties suggests the participation not of the mitochondrial genome but, rather, the nuclear genome for the perpetuation of plasmid in progeny varieties.     

Identification of genes suitable for DNA barcoding of morphologically indistinguishable Korean Halichondriidae sponges

The development of suitable genetic markers would be useful for defining species and delineating the species boundaries of morphologically indistinguishable sponges. In this study, genetic variation in the sequences of nuclear rDNA and the mitochondrial cytochrome c oxidase subunit 1 and 3 (CO1 and CO3) regions were compared in morphologically indistinguishable Korean Halichondriidae sponges in order to determine the most suitable species-specific molecular marker region. The maximal congeneric nucleotide divergences of Halichondriidae sponges in CO1 and CO3 are similar to those found among anthozoan cnidarians, but they are 2- to 8-fold lower than those found among genera of other triploblastic metazoans. Ribosomal internal transcribed spacer regions (ITS: ITS1 + ITS2) showed higher congeneric variation (17.28% in ITS1 and 10.29% in ITS2) than those of CO1 and CO3. Use of the guidelines for species thresholds suggested in the recent literature indicates that the mtDNA regions are not appropriate for use as species-specific DNA markers for the Halichondriidae sponges, whereas the rDNA ITS regions are suitable because ITS exhibits a low level of intraspecific variation and a relatively high level of interspecific variation. In addition, to test the reliability of the ITS regions for identifying Halichondriidae sponges by PCR, a species-specific multiplex PCR primer set was developed.     

丽蝇蛹集金小蜂几丁质酶基因鉴定与表达分析

几丁质酶是降解几丁质的糖苷水解酶,参与昆虫蜕皮、器官发育、免疫等重要生理过程。目前,寄生蜂等膜翅目昆虫中几丁质酶的鉴定以及功能研究仍较少。本研究基于生物信息学分析,在丽蝇蛹集金小蜂Nasonia vitripennis基因组中鉴定到14个几丁质酶基因,氨基酸个数介于312~2 682之间。系统进化分析表明丽蝇蛹集金小蜂几丁质酶分为9个亚家族,其中Group Ⅳ、Ⅶ亚家族可能通过基因串联复制而发生基因家族扩增。qRT-PCR分析结果表明几丁质酶基因的表达具有多样性,其中NvCht1、NvCht5、NvCht6、NvCht7 4个基因在1.5 d幼虫表达量最高,NvCht3在5 d幼虫表达量最高,NvCht8在幼虫期高表达。此外,幼虫组织中的表达分析结果表明NvCht4、NvCht5、NvCht10、NvCht12在表皮中高表达,NvCht7、NvCht8、NvCht13在肠道中高表达,NvCht9在脂肪体和表皮中高表达,NvCht11在唾液腺中高表达。本研究为寄生蜂几丁质酶的进化分析以及功能研究提供理论基础。     

Identification and expression analysis of mical family genes in zebrafish

Mical(molecule interacting with CasL)represent a conserved family of cytosolic multidomain proteins that has been shown to be associated with a variety of cellular processes,including axon guidance,cell movement,cell-cell junction formation,vesicle trafficking and cancer cell metastasis.However,the expression and function of these genes during embryonic development have not been comprehensively characterized,especially in vertebrate species,although some limited in vivo studies have been carried out in neural and musculature systems of Drosophila and in neural systems of vertebrates.So far,no mica/family homologs have been reported in zebrafish,an ideal vertebrate model for the study of developmental processes.Here we report eight homologs of m/ca/family genes in zebrafish and their expression profiles during embryonic development.Consistent with the findings in Drosophila and mammals,most zebrafish mical family genes display expression in neural and musculature systems.In addition,five mica/homologs are detected in heart,and one,micall2a,in blood vessels.Our data established an important basis for further functional studies of mica/family genes in zebrafish,and suggest a possible role for mica/genes in cardiovascular development.     

低温胁迫下6种木兰科植物的生理响应及抗寒相关基因差异表达

分析了6种木兰科植物对低温胁迫的生理响应及耐寒相关调控基因HSP90和WRKY33的差异表达,为木兰科植物抗寒机理的研究和抗寒品种的选育提供理论基础。结果表明,6种木兰科植物的低温LT50在-10.64—-22.06℃,从高到低依次为红花深山含笑、峨眉含笑、杂交含笑、阔瓣含笑、六瓣含笑和乐东拟单性木兰;低温过程中,6种木兰科植物叶片可溶性蛋白(SP)、游离脯氨酸(Pro)含量、超氧化物歧化酶(SOD)和过氧化物歧化酶(POD)活性呈先升高后降低的趋势,可溶性糖(SS)和丙二醛含量(MDA)则不断积累;筛选出REC、MDA、SP、SS和Pro作为6种木兰科植物抗寒性评价的关键指标;聚类分析将6种木兰科植物在抗寒性能上分为强、中、弱三类,分别为乐东拟单性木兰和六瓣含笑,阔瓣含笑、杂交含笑和峨眉含笑,以及红花深山含笑。对HSP90、WRKY33基因的差异表达分析表明,2个基因在6种木兰科植物中的相对表达量呈先升高后降低的趋势,在临近各树种LT 50时,2个基因的表达被强烈抑制且后期表达量不可逆。0℃时,2个基因的表达量差异不显著;-5℃时,2个基因开始被激活,表达量增加;-10℃时,HSP90、WRKY33基因在红花深山含笑叶片中的表达量较-5℃时分别下调了0.76倍和0.68倍,而在其他5个树种中的表达被进一步激活;-15℃时,HSP90和WRKY33基因在抗寒性中等的阔瓣含笑、杂交含笑、峨眉含笑中亦被强烈抑制,较-10℃时分别下调了0.38倍、0.33倍、0.32倍和0.71倍、0.72倍、0.74倍,在抗寒性强的乐东拟单性木兰和六瓣含笑中的表达被进一步激活;-20℃以后,2个基因在6个树种中的表达均被强烈抑制,但在抗寒性最强的乐东拟单性木兰中的表达量仍高于其他5个树种。抗寒基因的激活与表达是影响植物抗寒性的重要因素,抗寒性不同的树种对低温的应答机制明显不同。抗寒性越强的树种越能快速启动低温应答机制,激活抗寒相关基因的表达,进而调整生理生化活动以抵御和适应冷应力。不抗寒树种中抗寒基因的表达则受到抑制,降低了其对低温逆境的耐受能力。     

Screening of optimal reference genes for qRT-PCR and preliminary exploration of cold resistance mechanisms in Prunus mume and Prunus sibirica varieties

Molecular Biology Reports - Prunus sibirica and Prunus mume are closely related plant species that differ in cold tolerance. Hybrids of P. sibirica and true mume, belonging to the apricot mei...     

Identification and characteristics analysis of toll-like receptors family genes in yak

Toll-like receptors (TLRs) are innate pattern recognition receptors that play an important role in host resistance to pathogenic microbes. In this study, we cloned the coding region of the yak TLR family (1–10) genes and used bioinformatics to analyze gene characteristics. Real-time fluorescence quantitative polymerase chain reaction was used to detect TLR expression levels in different tissues. Yak TLR genes exhibited high homologies with other species. At the nucleotide level, yak shared more than 96 % homology with cattle and sheep and 75–87 % homology with human and mouse. At the amino acid level, yak shared 90–99 % homology with cattle and sheep and 64–86 % homology with human and mouse. Yak showed close evolutionary relationship with cattle and sheep, which formed a branch of mammals together with TLRs from human, horse, and mouse, among others, and formed a branch with a longer genetic distance with chicken. TLR1, 2, 6, and 10 and TLR7, 8, and 9 were clustered in 2 individual branches, respectively. Fluorescent quantitation results showed that TLRs were expressed in all yak tissues, but different members showed different expression patterns. TLR2, 4, and 6 showed the highest expression in the spleen, followed by ovary, small intestine, kidney, and liver. TLR1, 5, 7, 8, 9, and 10 were most highly expressed in the kidney and showed higher expression in the liver, kidney, spleen, and other tissues. Our results will be useful for studies on immune molecular mechanisms and disease resistance breeding of yak and other plateau animals.